[Isolation of platelet integrin GPIIbIIIa, preparation of its chymotrypsin fragments and IIIa subunit].
The method for isolation of glycoprotein IIbIIIa by chromatography on Concanavalin A-Sepharose and DEAE-Sephacel columns have been proposed. The 66 kDa, 27 kDa and 6 kDa fragments were produced by chymotrypsin digestion of GpIIbIIIa and purified with FPLC system on Superose-12 column under non denaturating conditions. N-terminal sequence analysis of fragments revealed that 66 kDa fragment contained NH2-terminal region, disulphide rich node, transmembrane and cytoplasmatic regions of IIIa subunit. 27 kDa fragment was a part of Ca(2+)-binding site of IIb subunit. 6 kDa fragment was shown to be from transmembrane and cytoplasmatic regions of IIb subunit. GpIIIa was isolated by gel-filtration on Sephacryl S-300 column of the GpIIbIIIa dissociated.
Iakubenko
V P
VP
Makogonenko
S M
SM
D000596:Amino Acids / Q000032:analysis; D015703:Antigens, CD / Q000737:chemistry / Q000302:isolation & purification*; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D003593:Cytoplasm / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006868:Hydrolysis; D039661:Integrin beta3; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000302:isolation & purification*; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*
D000596:Amino Acids; D015703:Antigens, CD; D039661:Integrin beta3; D010446:Peptide Fragments; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; D002918:Chymotrypsin; D002118:Calcium
Vidilennia trombotsitarnogo integrynu GPIIbIIIa ta otrimannia łogo khimotrypsinovykh fragmentiv i cubodinitsi IIIa